PHSL233 27th July 2023

L.6 Membrane Potentials: Diffusion, GHK equation

Slide 1: Recap from yesterdays lecture

Recap from yesterdays lecture:
 Yesterday left with chloride doesn’t contribute to the membrane potential
 The intracellular conc of chloride is determined by the extracellular concentration and the
membrane potential
 The reason for this
o Here is cell with diff concentrations across membrane
o Inside is -82mv
o Ran Nernst equation and tells us what happens to the ions or one particular ion in
the conditions that we have got
o Extracellular conc of sodium for example
 Plug numbers into Nernst equation and that says if the outside has 150 and
the inside is 10 which is a membrane potential that is the same as the
Nernst potential of 72mv, that is huge driving force
 It is how we can work out what the driving force for the ions is going to be,
which way they are going to move and how much they are going to move
 Sodium is a positively charged ion, its number is negative, we are going what
is the diff between 72 and -82 and the diff is 154, has a negative on it so
positive ions will enter the cell, we already know this, concentration
gradient for sodium to enter the cell, we know there is an electrical gradient
for sodium to enter the cell so sodium is always going to enter the cell, the
driving force is large
 -154 tells us that it will enter the cell (influx)
o Can do the same for potassium, again same thing, equilibrium potential of -90
 Reminder if you see potassium with 92 or 97 just a slightly diff but same
thing
 8 flux, don’t need a massive driving force for potassium to leave the cell,
have way more permeability to potassium so don’t need the massive driving
force, the gradient tells us essentially how much is going to go across but we
don’t need the massive driving force for it to leave, it will leave if there are
potassium channels open, to counter this all we have the sodium potassium
ATPase which is constantly recycling the potassium back into the cell as
potassium will always leave the cell and constantly pumping sodium out of
the cell bc if sodium can get into the cell via a channel
o Chloride for example
 Chloride sits at exactly -92 so CL is not going anywhere as the Nernst
potential matches the membrane potential and at that point there is not net
flux of ions, defiantly ions coming in and out of the cell but the number
leaving the cell is the same as the number entering the cell so there is no net
flux, so no driving force as it is in its happy place, in equilibrium
 So why do we have the situation where chloride is being bullied around, why do we have the
situation where it is the extracellular concentration of chloride and the membrane potential
that determines the intracellular concentration?
o reason is we don’t have a lot of permeability to chloride, we also have to look at the
Nernst equation when we look at driving forces
o Ie. extracellular chloride is 110, if increase the extracellular conc to 120, that shifts
the Nernst potential but again cause of faradays constant and other things it is not 1
 for 1, to get back to -82 will give us an Nernst potential to about -83/84 not a big
shift in number, to get but to -82 the intracellular chloride conc only has to shift to
5.4, it is the ratio of these things that matter not the absolute numbers, not a big
shift to change the intracellular conc by half a million then restarts that ratio, it is
easier to shift chloride around as it is sitting close to equilibrium potential, close to
membrane potential whereas potassium and sodium it is different and it comes
down largely to the permeability only need to open a few chloride channels to
reassert this balance across the membrane that brings is back to Nernst for chloride
for being the same for the membrane potential, we can drive it easily
 So sodium diffuses into the cell
 Potassium diffuses out of the cell
 The sodium potassium ATPase maintains the composition constant by
pumping stuff out
o Therefore we do not have equilibrium chloride in this case is so no net movement of
charge but sodium has a driving force and so does potassium so leave the cell so we
don’t have equilibrium, if cells get to equilibrium you will die as we need to move
things in and out
 The reason for membrane potential is that we want a gradient for sodium to enter so that
we can couple things to sodium to get them in cell
o So the membrane potential is maintained by the potassium gradient and sodium
gradient utilizes that gradient to drive diff transport protein configurations to move
things in and out of the cells to regulate lots of thing s ie. size of cell with osmolarity,
ie. cell increase osmolarity we shift some ions ot reduce osmolarity to stop the cell
increases too much in size
o pH is the same, pump hydrogen ions out in exchange for sodium ions, important for
health of cell, the sodium gradient is important so maintain the resting membrane
potential has to be way negative, the big it is the more efficient sodium is at moving
things around
o So we don’t have an equilibrium we have a stead state so what is a steady state
 Equilibrium is when no energy is required to keep the box at the same spot
on the table
 If tilt the box, lift one end up so at angle this box requires him to hold if for it
to stay there that is a steady state, as long as he is hold the boxy up there it
is happy but it requires some form of work or energy and in this case this
energy is coming from him holding the box so this is steady state
 Equilibrium no energy or movement transfer
 Just like when chloride is at equilibrium there is no net movement of
chloride ions, net flux 0
 Only steady state that we can maintain these gradients when cells stead
state, work is don’t by the sodium potassium ATPase, if this gets blocked or
something stops working everything else gets run down, it will mean
potassium and sodium will find an equilibrium and when this is done the cell
stops doing things and the cells die
 Remember this is talking about an individual cell and we are made up of
millions of cells so all of these things working together
 
Relationship between Ex and Vm
We are interested in the theory of what is going on with cells, they just do it, we try to deconstruct
how they are doing it
 What is the relationship between Ex and Vm
 Ex is the Ernest potential or the equilibrium potential and Vm is the membrane potential
  Ex calculates the diffusion potential for one ion
 Ex does not calc the membrane potential (Vm) bc we know the Vm is made up of all the
contributions of all of the ions, most of the ions
 Key thing is only ions that can cross the membrane contribute to the membrane potential, if
you don’t have a mechanisms for cross the membrane ie. channel or transporter, you as an
ion don’t contribute to the membrane potential and that is why when we say potassium
channels that have way more in the cell, the permb is 10,000x mores than it is for sodium,
that is why the membrane potential sits close to the membrane potential for potassium and
not sodium
 The key thing the Ex misses is that it is only accounting for one ion and does not count for
any permeability it is an ideal, an ideal sitting there, so if the membrane happens to move
the EX potential for potassium you will get no movement of potassium bc we calculated
that, potassium ions don’t know what's going on they cant do math's so permeability is key if
you want to know what is going on with the membrane potential. That is why the Ex tells us
where the happy place for ion would be and if the membrane potential ever gets there it
would stop the movement of ions
 Equation top right, potassium (RT/zF can shift, R is gas constant, z is constants, F is a
constant but temperature T can change it!) - we will do this calc in the lab so important to
know, therefore the diff between the Ex potential for potassium and the membrane
potential equals the diving force for the efflux of potassium
 If the ratio changes, the ratio of ions across the membrane then the Ex will shift, if we shift
the ratio has changed, the Ex would change to -82 and if that happens the potassium will be
at equilibrium, the membrane potential won't be but potassium will be so there will be no
net movement of potassium ions, simply by moving 2 mmolar of potassium from outside the
cell t inside the cell we can reach equilibrium for potassium and we don’t want that!
 Sodium potassium ATPase is doing all it can to make sure we don’t shift 2mmoler of
potassium into the cell
 You could so this artificially by injecting potassium chloride into some's blood stream, with
the blood that has a high level of potassium when it gets to the heart extracellular fluid vs
intracellular fluid you get a depolarization, massive one and that would cause you to have a
massive heart attack
 8 bananas, took potassium from that and injected that would kill them
 Even eating 8 bananas is risky
 
Relationship between Ex, [X]o, and [X]i
So what if we change the concentrations
 If the chemical gradient is reduced, the electrical gradient gets smaller as just a reflection of
the chemical gradient, so therefore the Ek gets closer to zero
 If the concentration is the same across the membrane, there will be no electrical driving
force
 Remember the close the top number and the bottom number are together, the small that
number will be, weather it is a positive or negative it will move closer to zero
 
 If the chemical gradient is increased, we get a bigger number, as it is potassium it will be a
bigger number as high conc on the inside not the outside, bigger number so a bigger driving
force
 So simply by changing the ratio across the membrane we alter the membrane potential,
 
 it is about the ratio not the absolute numbers
  So if they are both altered but the ratio is the same, the ratio to inside and outside is the
same as it would be so if you do the Ex equation = exactly the same membrane potential -
91mv
 Highlights defiantly the ratios that are important
 
 If you swap the inside to the outside conc, you get a positive number! Ratios!
 That is why sodium is a positive high conc outside and low inside so that’s why potassium is
a big negative number as big conc on the inside and a small conc on the outside, it is just the
flipping of that ratio that makes a diff
 
Membrane Potential (Vm)
 Membrane potential is a combination of diffusion potentials
 What is a diffusion potential? It is the same as the equilibrium potential and same as the Ex
potential and the same as reversal potential (another name - NEED TO KNOW AS THEY MAY
TRY TO CONFUSE8 YOU)
 Potassium drives the membrane potential towards its happy place as it is very permeable
 Sodium would like to drive the membrane potential towards its happy place, there is sodium
permeability but not as much as potassium permeability
 Often potassium permeability is known as the leak current bc it facilities this drive towards -
90mv
 Both contribute but the contribution depends on their relative permeability
 Membrane potential is less than -90mv, textbooks will say for a given cell, it may be
anywhere between -60 and -90, simply comes down to what is the permeability to
potassium and what is permeability to sodium
 Potassium wants to drive it way negative, sodium wants positive so they find a happy
medium based on permeability
 That brings to equation, this is Goldman equation
 
Vm described by Goldman equation
 This is simply the permeability of potassium x the conc of potassium outside dived by the
permeability of potassium which is the same no, x the conc of potassium on the inside, this
is just a glorified Ex equation, it include all of the ions we are interested in
 Some people would include calcium here ie. in the brain but we are doing the generic one,
we are doing epithelial physiology here and cell transport
 So typically we just have potassium sodium and chloride
 Why chloride in there bc it can do stuff
 Just a note the permeability of sodium section the same as potassium
 Then we get to chloride and we invert it bc all of the ions are univalent so Z is excluded, it is
emitted bc it is always 1 for these guys but this is the only negative ion so to incorporate it
correctly in the equation we have to evert it
 
 Bc the membrane is more permeable to potassium than sodium the membrane potential
approached the equilibrium potential for potassium towards -90mv
 The gradients for sodium and potassium determine the membrane potential if chloride is at
equilibrium, why is this relevant? Bc in a couple lectures we will talk about what happens in
epithelial cells when chloride is not at equilibrium
 Vm membrane potential is not an equilibrium potential it is a steady state potential
o It is a steady state potential bc it requires sodium and potassium to stay away from
their equilibriums
 
Relative Permeability
Permeability
 It is difficult to measure absolute permeability, permeability is experimentally determined!!
 You will always be given the permeability
 Basically work on the assumption that most cells have about the same permeability bc most
of the time that is true
 So typically we talk about the relative permeability, the ratio
 P is generally expresses as a ratio denoted as on the bottom
o So this says that K is 100% relatively high permeability to potassium compared to
sodium that has much less 0.05 and chloride sits in the middle
o Relative permeability
o So know this particular cell will be more permeably to potassium probably bc more
channels and higher open time in comparison to sodium and chloride in the middle
bc it gets boss around
 
Question: What drives changes in Vm?
Interactive question
 This is what a AP looks like
 Can look at sodium and potassium
 So at resting membrane potential, permeability for potassium is lots compared to sodium
which is a little bit permeability
 When membrane potential suddenly becomes way more positive, why bc three Pk all of a
sudden the permeability of sodium has gone up to maybe 5 sodium, reason membrane
potential goes up is bc massively increase permeability to sodium bc we open a lot of voltage
gated sodium channels
 Then peak
 Then membrane potential is dropping so now have sodium permeability dropping as sodium
channels are closing and we have opens a whole bunch of potassium channel and that drives
back down
 Next get the hyper-polarization we get close to Ek as we have even more potassium
channels open, these are transient potassium channels as they will open and then close after
a finality amount of time so sodium is dropping back
 Then get back to re-establish resting membrane potential where potassium channels are
closing back to three arrows and back to only 1 sodium
 Permeability changes as ions open
 
 
 
Value of an equilibrium potential
What is the value of an equilibrium potential
 This is an entirely theoretical proposal
 Have membrane potential down at -100 up to positive 65
 Looking at the relative permeability to the different ions, essentially are the channels there
and are they open, could be channels there that is closed
 So relative permeability if we shut down any sodium ion channels, got rid of any chloride
channels and just had potassium ion channels then the membrane potential would as we
know match the Ex, the membrane would shift its potential so -97 (don’t sweat slight diff)
 
 Then if we got ride of all potassium channels, no sodium channels so only permeability to
chloride so chloride permeability is 1 as all relative then membrane potential would match
the Ex potential for chloride
 
 Same for sodium, close all of the other channels for potassium and chloride we would get
the membrane potential shift to, the Ex potential, if we kept this state here the membrane
potential would just stay here and sit at positive 61 until did something to alter the
permeability of other ions
 
 Rest shows normal, it is why relative permeability maintains the resting potential at the
umber it is maintained at if you shift those permeability we shift what that number is at
 
Goldman-Hodgkin-Katz equation in action
Math's of it
 Need this for the lab
 Have to do it in order
 R, T and F given to you, only diff is if the temperature changed!!
 Key thing is multiply each
 Permeability is 1
 Conc outside is 5
 Again given numbers
 So break it down
Add together
 When get down to Vm - 0.0254 Ln (0.0805)
 Work it backwards go 0.0805 natural log = -2.519
 Then multiple that by the constant
 Givers voltage so never report in volts!!!
 Milli volts so have to convert, it is a rule!!
 Exam question would never get full make if that was answer
 So convert by 1000x (1000 mili volts in a volt)
Remember concentration and permeability that is all theory, need to know and understand
Permeability is given but need to understand bc we have to do other things
 
Permeability, Ohm's law, & conductance
 Permeability is the capability of an on to flow across the membrane (weather they are
actually moving or not)
 We don’t talk about permeability as a thing we talk about conductance
 Conductance is what measure the movement of charge across the membrane, actual
measurement of charge across the membrane
  Takes us back to AP slide, conductance is the measurement of ions going across the
membrane, it is the practical side of the theory
 This is related to Ohms law = V=IR, voltage is equal to currant x resistance
 Volts, amps, resistance which is Ohms
 
Ohm's law in physiology
 Flux of ions passing through a channel, that generates a currant i , this is influence by the
electrical field across the membrane we know that the movement of ions can be altered by
the membrane potential the two are related, the resistance of that flux through the ions
channel, resistance resist the flow through the current
 That mean if you have a membrane shifting over, and no channels for potassium it has a high
resistance to potassium,
 If you insert channels the resistance drops bc now providing a mechanisms for potassium to
move
 We tend to think about it in terms of current as we measure the currant, we don’t measure
the permeability we just calc I theoretically, we actually measure the currant
 We look at the resistance
 The current tells us ions are flowing, tells us how many, small currant not many ions are
flowing, larger current lots are flowing, inversely proportional, the bigger the resistance, the
smaller the current will be, the resistance comes down to essentially the permeability, how
many channels are in the membrane, how many open, how much they let though
 Voltage is proportional to current so big voltage = big potential diff across the membrane so
have permeability across the membrane we get a large flow of ions across membrane, this
related back to the theory we have talked about
 Resistance is in the inverse, lots of resistance so smaller current ie. might have channels in
the membrane but don’t have many open, if we open the channels then we reduce the
resistance
 A large potential diff (v) across the membrane results in larger current flow
 In a wire, current is carried by electrons in biological solution it is by the movement of ions,
positively and negatively charged
 
Conductance
Conductance
 Given symbol G, units is Siemens, often used instead of current i
 Don’t often talk about the current we use conductance more often in the practicalities of
what we talk about
 There are for example potassium channels known as big K that allow lot of conductance,
when those channels are open they conduct a lot of potassium ions, medium Ks, medium
conductance potassium channels, when open only medium level of conductance relative to
big K or small K, there are over 80 potassium channels each with diff characteristics
 Essentially how easy does ion X flow through the membrane and be influenced by all the
other things so the greater the conductance of an ion, the more it will influence the
membrane potential, shift the idea of permeability and conc gradients which still exist but
for practical reason we talk about the conductance of an ion
 Bigger conductance more ions will flow
 Will influence the membrane bc if we talk about potassium, cells have high poasttium8
conductance (high permeability, high conductance) so means potassium will dominate, we
can change the conductance like the figure earlier, the conductance go up for the ions as
they contribute more and more to the membrane potential
 Conductance is G, it ignores voltage but talks about resistance
 If resistance is small the conductance will be high and vie versa
  That is practice of all theory